Effects of media and promoters on different lipid peroxidation assays in stallion sperm
Effects of media and promoters on different lipid peroxidation assays in stallion sperm
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DOI：
10.1016/j.anireprosci.2019.106199
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发表时间：
2019-12-01
影响因子：
2.2
通讯作者：
Varner, Dickson D.
中科院分区：
文献类型：
--
作者：
Ghosh, Sharmila;Serafini, Rosanna;Varner, Dickson D.
不同的培养基和促进剂对活种马精子脂质过氧化（LPO）的影响尚未报道。本研究的目的是使用四种不同的流式细胞术测定（即，BODIPY、Liperfluo、4-羟基壬烯醛[4 HNE]、丙二醛[MDA]）。使用Liperfluo、4 HNE和MDA测定检测到显著的培养基X启动子相互作用（P < 0.05）;因此，数据按培养基和启动子分类。在纳入乳基培养基的情况下，使用所有LPO测定的对照样品中的LPO浓度相似。铁作为LPO产生的促进剂，其作用是介质依赖性的，并且当对所有测定进行评估时，基于牛奶的介质保护精子免受铁诱导的LPO产生。相比之下，当在除BODIPY之外的所有试验中使用时，铁促进了BWW中稀释的精子中的LPO，这可能是因为使用该试验的靶分子不同。紫外线是最有效的LPO启动子，所有的媒体和测定评价。数据表明，在LPO产生途径的早期（基于BODIPY和Liperfluo测定），乳基补充剂通常比BWW更具LPO保护性，但在LPO产生的后期阶段（基于4 HNE和MDA测定），其保护性较低。使用不同的媒体和启动子的LPO允许测定早期和晚期阶段的LPO在可行的种马精子。
Effects of different media and promoters on lipid peroxidation (LPO) in viable stallion sperm have not been reported. Aims of this study were to determine effects of three media (INRA-96 (TM), Equipro CoolGuard (TM), and Biggers, Whitten and Whittingham [BWW]), and promoters (iron sulfate-Fe; ultraviolet light-UV; or control-no exposure to promoters) on viable sperm LPO using four different flow cytometric assays (i.e., BODIPY, Liperfluo, 4-hydroxylnonenal [4HNE], malonaldehyde [MDA]). Significant media x promoter interactions were detected using the Liperfluo, 4HNE, and MDA assays (P < 0.05); therefore, data were sorted by media and by promoters. With inclusion of milk-based media, there were similar concentrations of LPO in control samples with use of all LPO assays. The effect of iron, as a promoter of LPO production, was media dependent, and milk-based media protected sperm from iron-induced LPO production when there were assessments with all assays. In contrast, iron promoted LPO in sperm diluted in BWW when there was use of in all assays, except BODIPY, probably because of the different target molecule with use of this assay. Ultraviolet light was the most potent LPO promoter with all media and assays evaluated. Data indicate milk-based extenders are generally more LPOprotective than BWW early in the LPO production pathway (based on BODIPY and Liperfluo assays), but are less protective during the later stages of LPO production (based on 4HNE and MDA assays). The use of different media and promoters of LPO allowed for determination of early and late stages of LPO in viable stallion sperm.